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Immune system of the bee and relation with CCD

Cardinium is a bacterial symbiont infecting many species of arthropods, and is associated with manipulation of host reproduction. Cardinium is the causal agent of asexual reproduction, or thelytoky, in the chalcidoid parasitoid wasp Encarsia hispida. Feeding antibiotics to the infected adult females results in uninfected male offspring. Here, we show that these males are diploid. Diploid males are extremely unusual in the large hymenopteran superfamily Chalcidoidea, and, to our knowledge, have never before been associated with symbiont infection in this group. These findings indicate that at least in E. hispida, diploidy restoration is a necessary condition but not sufficient to elicit female development. Cardinium is required to feminize diploid male embryos and thus must interact with elements of the host sex determination system. In addition, our data suggest that Cardinium is necessary for the fertility of E. hispida; antibiotic curing of Cardinium reduces offspring production of adult females

A recent review of karyology of the family Eulophidae showed that many genera remain totally untouched by chromosomal investigation, among which the genus Thripobius. Karyotype analysis was carried out on Thripobius javae by a technique for single individuals. The diploid karyotype of T. javae consists of two pairs of large metacentric chromosomes, three pairs of shorter metacentrics and a pair of very small acrocentric chromosomes, thus showing 2n=12 and NF=22. The set of the haploid karyotype with five long bi-armed chromosomes and a short acrocentric corresponds to that already reported for the Eulophidae family.

A simple and rapid method for the extraction of total nucleic acid from honeybee and mite, useful either as template for RT-PCR or in nucleic acids hybridization, was developed. Sensitivity of the methods were evaluated up to 10(9) and 10(6) dilution of TNAs extracted from a single honeybee, for reverse transcriptase polymerase chain reaction and molecular hybridization respectively. The two diagnostic methods developed could be useful for the study of the molecular biology and the pathology of DWV. For practical applications dot-blot hybridization could be used in order to study the incidence of DWV in honeybees populations. The method is enough sensitive, rapid and less affected by contamination problems compared to RT-PCR and thus it could be applied to the sanitary certification of honeybees and their products

Mites in the genus Varroa are obligate ectoparasites of honey bee populations worldwide. Recent evidence from morphological, geographical, and especially genetic variation has spurred an important revision of Varroa taxonomy. Specifically, mitochondrial DNA (mtDNA) evidence suggests that the main mite pest on western honey bees (Apis mellifera) is not Varroa jacobsoni, as first described, but a distinct species now named Varroa destructor. Genetic markers also have been used to support a taxonomic basis for regional differences in how Varroa mites impact honey bees. Recent morphometric and molecular studies confirmed the presence of the species V. destructor also in the apiaries of the Campania region of southern Italy. In the three-year period 2001-2003 a survey was conducted in 118 municipalities of the five provinces of the Campania region in order to add data to the limited epidemiological information available regarding Varroa destructor in this zone. The level of infestation by the mite was assessed on a total of 521 apiaries (241 apiaries were inspected on 2001, 154 on 2002, and 126 on 2003). In each apiary, 100 comb cells were examined and in each province the level of infestation was calculated using the following formula: (number of Varroa specimens/number of open comb cells) x 100. In order to display the level of infestation, Geographical Information Systems were used in order to draw parasitological maps

The Red Palm Weevil (RPW) is one of the major pests of palms, frequently leading to the plants death. Action plans and the development of bio/physical strategies to contrast RPW diffusions are strongly recommended due to the serious concerns related to environmental pollution and insects’ resistance to chemicals. In the present study, we investigated morphological alterations of the ovaries and testes in adult RPW exposed to 2.45 GHz for 5, 15, and 30 s. During these treatments, the relative increase in temperature and the days of survival after irradiation were monitored. Then, RPWs were processed for macroscopical and microscopical analysis. Histological lesions of the ovaries and testes were characterized by the degeneration and necrosis of germinal cells, which increased with the increase in the time of irradiation and the temperature. By the same token, an increase in the temperature of irradiated insects was associated with a decrease in their survival time. These observations lead us to conclude that MWs could represent a useful tool for reducing or eliminating the reproductive capacity of this dreaded insect.

The neonicotinoid Clothianidin has a negative impact on NF-κB signaling and on immune responses controlled by this transcription factor, which can boost the proliferation of honey bee parasites and pathogens. This effect has been well documented for the replication of deformed wing virus (DWV) induced by Clothianidin in honey bees bearing an asymptomatic infection. Here, we conduct infestation experiments of treated bees to show that the immune-suppression exerted by Clothianidin is associated with an enhanced fertility of the parasitic mite Varroa destructor, as a possible consequence of a higher feeding efficiency. A conceptual model is proposed to describe the synergistic interactions among different stress agents acting on honey bees., Pesticides could increase bees’ susceptibility to parasites, but the nature of this interaction has been unclear. Here the authors show that the pesticide Clothianidin reduces the wound healing immune response in bees, allowing the ectoparasitic Varroa mites to consume more bee hemolymph and amplify reproduction.

The association between the deformed wing virus and the parasitic mite Varroa destructor has been identified as a major cause of worldwide honeybee colony losses. The mite acts as a vector of the viral pathogen and can trigger its replication in infected bees. However, the mechanistic details underlying this tripartite interaction are still poorly defined, and, particularly, the causes of viral proliferation in mite-infested bees. Here, we develop and test a novel hypothesis that mite feeding destabilizes viral immune control through the removal of both virus and immune effectors, triggering uncontrolled viral replication. Our hypothesis is grounded on the predator–prey theory developed by Volterra, which predicts prey proliferation when both predators and preys are constantly removed from the system. Consistent with this hypothesis, we show that the experimental removal of increasing volumes of haemolymph from individual bees results in increasing viral densities. By contrast, we do not find consistent support for alternative proposed mechanisms of viral expansion via mite immune suppression or within-host viral evolution. Our results suggest that haemolymph removal plays an important role in the enhanced pathogen virulence observed in the presence of feeding Varroa mites. Overall, these results provide a new model for the mechanisms driving pathogen–parasite interactions in bees, which ultimately underpin honeybee health decline and colony losses.

Clothianidin is a widely used neonicotinoid insecticide, which is a potent agonist of the nicotinic acetylcholine receptor in insects. This neurotoxic compound has a negative impact on insect immunity, as it down-regulates the activation of the transcription factor NF-κB. Given the evolutionary conserved role of NF-κB in the modulation of the immune response in the animal kingdom, here we want to assess any effect of Clothianidin on vertebrate defense barriers. In presence of this neonicotinoid insecticide, a pro-inflammatory challenge with LPS on the human monocytic cell line THP-1 results both in a reduced production of the cytokine TNF-α and in a down-regulation of a reporter gene under control of NF-κB promoter. This finding is corroborated by a significant impact of Clothianidin on the transcription levels of different immune genes, characterized by a core disruption of TRAF4 and TRAF6 that negatively influences NF-κB signaling. Moreover, exposure to Clothianidin concurrently induces a remarkable up-regulation of NGFR, which supports the occurrence of functional ties between the immune and nervous systems. These results suggest a potential risk of immunotoxicity that neonicotinoids may have on vertebrates, which needs to be carefully assessed at the organism level.

The invasive Red Palm Weevil is the major pest of palms. Several control methods have been applied, however concern is raised regarding the treatments that can cause significant environmental pollution. In this context the use of microwaves is particularly attractive. Microwave heating applications are increasingly proposed in the management of a wide range of agricultural and wood pests, exploiting the thermal death induced in the insects that have a thermal tolerance lower than that of the host matrices. This paper describes research aiming to combat the Red Palm pest using microwave heating systems. An electromagnetic-thermal model was developed to better control the temperature profile inside the palm tissues. In this process both electromagnetic and thermal parameters are involved, the latter being particularly critical depending on plant physiology. Their evaluation was carried out by fitting experimental data and the thermal model with few free parameters. The results obtained by the simplified model well match with both that of a commercial software 3D model and measurements on treated Phoenix canariensis palms with a ring microwave applicator. This work confirms that microwave heating is a promising, eco-compatible solution to fight the spread of weevil.

Flower pollen is collected by honeybee foragers, adhered on their rear legs and transported into the hives in the form of pellets. Once in the hives, bee pollen is moisturised with nectar and bee mouth secretions and due to enzymatically modifications it becomes the so-called bee-bread, the protein reservoir of young bees. Bee pollen can be artificially removed from bee legs and collected by using specific systems, the bee pollen traps. Bee pollen is commercialized for human consumption as fresh product and after freezing or drying. Although bee pollen is nowadays largely consumed in developed countries, as food or food supplement according to local legislation, little is known on its safety related to microbiological hazards. In this work, we aimed to characterize for the first time the microbiological profile of Italian bee pollen in fresh, frozen and dried form collected along an entire harvesting season. Moreover, monthly microbiological analyses were performed on frozen (storage at -18°C) and dried (storage at room temperature) bee pollen over a 4 months period. Further aim of this work was the evaluation of the possible impact on production level of three different traps used for pollen collection. Our results on microbial contamination of fresh and frozen bee pollen show that a more comprehensive microbiological risk assessment of bee pollen is required. On the other side, dried pollen showed very low microbial contamination and no pathogen survived after the drying process and during storage.

The red palm weevil (RPW), Rhynchophorous ferrugineus Oliver (Coleoptera: Curculionidae), is an economically important, tissue-boring pest of date palm in many parts of the world. The aim of the work was to investigate the effects of two plant oil-based products (BRK6 and BRK9) on the mortality of RPW (Exp. 1) and morpho-physiological characteristics of palm (Exp. 2). Laboratory experiments showed that the two mixture of oils BRK6 and BRK9 showed larvicide and adulticide properties. These properties were more pronounced on BRK9 than BRK6. The lethal concentration for 50% of individuals (LC50) calculated by probit, of BRK6 was 0.22% (for the 90% of individuals LC90 = 0.34) while for BRK9 the calculated LC50 was 0.10% (LC90 = 0.66). These data show that a smaller amount of BRK9 oil was required to induce the 50% of mortality of RPW larvae. As regards to the adults, BRK9 and BRK6 were effective only at the highest concentration (2 and 5%, respectively). Finally, injection of BRK9 into the palm trunk showed to promote chlorophyll content and photosynthesis of leaves thanks to a good translocation of zinc.

Honey bee colony losses are triggered by interacting stress factors consistently associated with high loads of parasites and/or pathogens. A wealth of biotic and abiotic stressors are involved in the induction of this complex multifactorial syndrome, with the parasitic mite Varroa destructor and the associated deformed wing virus (DWV) apparently playing key roles. The mechanistic basis underpinning this association and the evolutionary implications remain largely obscure. Here we narrow this research gap by demonstrating that DWV, vectored by the Varroa mite, adversely affects humoral and cellular immune responses by interfering with NF-κB signaling. This immunosuppressive effect of the viral pathogen enhances reproduction of the parasitic mite. Our experimental data uncover an unrecognized mutualistic symbiosis between Varroa and DWV, which perpetuates a loop of reciprocal stimulation with escalating negative effects on honey bee immunity and health. These results largely account for the remarkable importance of this mite–virus interaction in the induction of honey bee colony losses. The discovery of this mutualistic association and the elucidation of the underlying regulatory mechanisms sets the stage for a more insightful analysis of how synergistic stress factors contribute to colony collapse, and for the development of new strategies to alleviate this problem.

Honey bees (Apis mellifera L.) are bioindicators of environmental pollution levels. During their wide-ranging foraging activity, these hymenopterans are exposed to pollutants, thus becoming a useful tool to trace the environmental contaminants as heavy metals, pesticides, radionuclides and volatile organic compounds. In the present work we demonstrate that bees can also be used as active samplers of airborne particulate matter. Worker bees were collected from hives located in a polluted postmining area in South West Sardinia (Italy) that is also exposed to dust emissions from industrial plants. The area is included in an official list of sites of national interest for environmental remediation, and has been characterized for the effects of pollutants on the health of the resident population. The head, wings, hind legs and alimentary canal of the bees were investigated with Scanning Electron Microscopy coupled with X-ray spectroscopy (SEM-EDX). The analyses pointed to specific morphological and chemical features of the particulate, and resulted into the identification of three categories of particles: industry -, postmining -, and soil –derived. With the exception of the gut, all the analyzed body districts displayed inorganic particles, mostly concentrated in specific areas of the body (i.e. along the costal margin of the fore wings, the medial plane of the head, and the inner surface of the hind legs). The role of both past mining activities and the industrial activity close to the study area as sources of the particulate matter is also discussed. We conclude that honey bees are able to collect samples of the main airborne particles emitted from different sources, therefore could be an ideal tool for monitoring such a kind of pollutants.

The decline of honeybee colonies and their eventual collapse is a widespread phenomenon in the Northern hemisphere of the globe, which severely limits the beekeeping industry. This dramatic event is associated with an enhanced impact of parasites and pathogens on honeybees, which is indicative of reduced immunocompetence. The parasitic mite Varroa destructor and the vectored viral pathogens appear to play a key-role in the induction of this complex syndrome. In particular, the Deformed Wing Virus (DWV) is widespread and is now considered, along with Varroa, one of the major causes of bee colony losses. Several lines of evidence indicate that this mite/DWV association severely affects the immune system of honeybees and makes them more sensitive to the action of other stress factors. The molecular mechanisms underpinning these complex interactions are currently being investigated and the emerging information has allowed the development of a new functional model, describing how different stress factors may synergistically concur in the induction of bee immune alteration and health decline. This provides a new logical framework in which to interpret the proposed multifactorial origin of bee colony losses and sets the stage for a more comprehensive and integrated analysis of the effect that multiple stress agents may have on honeybees.

Rhynchophorus ferrugineus Oliv., known as the red palm weevil (RPW), has quickly spread in Southern Europe, infesting and destroying an increasing number of palms, particularly the Phoenix canariensis ones. Of the various techniques suggested for treating the palms, high power microwave applications are considered an attractive, eco-compatibile solution. However, in order to correctly design the exposure system, a knowledge of the electromagnetic properties of the materials involved is required. In this paper, we present a broad-band electromagnetic characterization in the 0.4–18 GHz frequency range of the tissues (both healthy and damaged) of the P. canariensis, with different moisture content, and of the R. ferrugineus in different stages (larva, pupa and adult). The palm tissues had a high water content and a dielectric model of the vegetation was applied to the experimental data in order to estimate the volume fraction of free water and of the bulk vegetation-bound water mixture as well as the...